苏丹草RAPD反应条件的优化与应用  

Optimization of RAPD Reaction System for Construction of the DNA Fingerprinting in Sudangrass

作　　者：李杰勤[1] 王丽华[2] 詹秋文[1] 李云飞[1] 

机构地区：[1]安徽科技学院,安徽凤阳233100 [2]四川农业大学水稻研究所,四川温江611130

出　　处：《中国农学通报》2006年第12期397-400,共4页Chinese Agricultural Science Bulletin

基　　金：安徽省科技攻关计划重点项目"饲草新品种选育及其在安徽草山草坡改良中的利用"(No:06013104B);安徽省高校学科拔尖人才基金项目:;安徽科技学院引进人才项目"高粱与苏丹草及其近缘种的系谱分析"(ZRC200564);安徽科技学院重点学科建设基金项目"苏丹草与高粱SSR和RFLP及其与杂种优势关系的研究"(No:YZD2004-03)

摘　　要：为研究不同因素对苏丹草随机扩增多态性DNA(RAPD)反应体系的影响,建立并优化反应体系;再利用优化体系绘制了DNA指纹图谱以区分两个高丹草品种。提取苏丹草的基因组DNA,分别设置Mg2+浓度为1.0mmol/L、1.5mmol/L、2.0mmol/L、2.5mmol/L、3.0mmol/L;退火温度为34℃、35℃、36℃、37℃、38℃;Taq酶含量为0.6U、0.8U、1.0U、1.2U、1.4U进行PCR扩增。结果表明:Mg2+2.0mmol/L,退火温度36度,Tap酶活性值1.2U是进行苏丹草RAPD分析的最优反应条件。利用最优反应条件绘制了皖草2号和皖草3号的DNA指纹图谱,在该图谱中有A、B两条特征带,可以区分皖草2号和皖草3号。To establish and optimize reaction system, the effects of different factors on random amplified polymorphic DNA （RAPD） reaction system in Sudangrass were studied and the fingerprinting of two hybrids to differentiate them were constructed. Genome DNA was isolated. The different concentration of Mg^2＋（1.0mmol/L, 1.5 retool/L, 2.0 mmol/L, 2.5 retool/L, 3.0 mmol/L）, annealing temperature （34℃, 35℃, 36℃, 37℃, 38℃） and Tap DNA polymerase activity （0.6U, 0.8U, 1.0U, 1.2U, 1.4U） were designed for PCR amplification. The results were as follow： the optimization of a 20μl; PCR system was coupled with the concentration of Mg^2＋ 2.0 mmol/L, Taq DNA polymerase activity 0.6U and the annealing temperature 36℃. The DNA fingerprinting of two hybrids was constructed by RAPD marker. Two special bands, A and B, can differentiate Wancao2 and Wancao3 in the fingerprinting.

关 键 词：苏丹草 RAPD 指纹图谱 优化 

分 类 号：S544.1[农业科学—作物学]